The preparation method of beauverum

A technology of beauveria and fusarium, applied in the field of industrial microorganisms, can solve the problems of unsuitability for industrial production, cumbersome operation steps, and few samples to be processed, and achieve the effect of long time-consuming change, strong controllability, and short process cycle

Active Publication Date: 2016-04-27
NCPC NEW DRUG RES & DEV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The three separation and purification methods mentioned above all have the same disadvantages: namely, the operation step

Method used

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  • The preparation method of beauverum
  • The preparation method of beauverum
  • The preparation method of beauverum

Examples

Experimental program
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Example Embodiment

[0030] Example 1

[0031] (a) Preparation of Fusarium LF061 seed solution

[0032] The seed culture medium was prepared according to the following ratio: YeastExtract2g, Peptone4g, glucose 4g, and distilled water to make the volume to 200ml; adjust the pH to 5.8, and sterilize at 121°C for 20min.

[0033] The Fusarium LF061 was inoculated into a seed medium, and cultured at 28°C at 120 rpm for 48 hours to obtain seed liquid.

[0034] (B) Fusarium LF061 seed liquid is fermented to prepare Beauverin:

[0035] Liquid fermentation medium preparation: glucose 5g, soluble starch 20g, industrial peptone 30g, potassium chloride 0.1g, potassium dihydrogen phosphate 3g, sodium nitrate 1g, magnesium sulfate heptahydrate 0.5g, calcium carbonate 1g, add water to make the volume to 1000ml, Natural pH value, sterilize at 121℃ for 30min;

[0036] The Fusarium LF061 seed solution was inoculated into the liquid fermentation medium at an inoculum of 15% by volume, and fermented in a shake flask. The ferme...

Example Embodiment

[0038] Example 2

[0039] (a) Preparation of Fusarium LF061 seed solution

[0040] Prepare seed culture medium according to the following ratio: YeastExtract 20g, Peptone 40g, glucose 40g, add distilled water to make the volume to 2000ml; adjust the pH value to 5.3, and sterilize at 121°C for 20 minutes.

[0041] The Fusarium LF061 was inoculated into a seed medium, and cultured at 30°C at 130 rpm for 72 hours to obtain a seed liquid.

[0042] (b) Fusarium LF061 seed liquid is fermented to prepare Beauverin

[0043] Liquid fermentation medium preparation: glucose 300g, soluble starch 900g, industrial peptone 1500g, potassium chloride 6g, potassium dihydrogen phosphate 150g, sodium nitrate 45g, magnesium sulfate heptahydrate 3g, calcium carbonate 60g, add water to make the volume to 30L, pH value Naturally, sterilize at 121℃ for 30min;

[0044] The Fusarium LF061 seed solution was inoculated into the liquid fermentation medium at an inoculum of 5% by volume, and fermented in a 50L fermen...

Example Embodiment

[0045] Example 3

[0046] Take 10L of the beauvericin fermentation broth prepared in Example 2, and the fermentation unit is 425 μg / mL. The beauverin fermentation broth was filtered to obtain 2.5 kg of mycelium. Add 5.0 L of methanol to the above mycelium, stir and extract for 4 hours and then filter to collect the beauvericin extract. The extract was concentrated under a vacuum of 0.09 MPa at 40°C until no liquid flowed out to obtain a crude extract of Beauvericin with a purity of 53% (see the attached liquid chromatogram) figure 1 ). Take 10g of the crude extract of Beauverin, dissolve it with 60mL of methanol, and add the sample to a medium pressure chromatography column (Ф5.5×50cm) of UniPS30-300 polymer microspheres with a volume of 1000mL, using 80% volume percent After washing 2000 mL with methanol aqueous solution as the mobile phase, it was washed with methanol and received in separate tubes at a flow rate of 1500 mL / h. HPLC detection showed that the eluates whose pu...

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Abstract

The invention discloses a preparation method of beauvericin. The method comprises the steps of 1a, preparation of a fusarium LF061 seed solution preparation:inoculating a slant culture or spore liquid of strain LF061 into a seed culture medium, and culturing for 48-72 hours with a rotating speed of 120-160rppm at 28-32 DEG C to obtain the seed solution; 1b, preparation of beauvericin from the fusarium LF061 seed solution through fermentation: inoculating the seed solution of which the inoculation amount is 5-15% in percentage by volume into a liquid fermentation culture medium, wherein the fermentation temperature is 28-32 DEG C, the rotating speed is 120-160rpm, the ventilation volume is 10-30L/min, and the fermentation time is 96-144 hours; separating and refining obtained fermentation liquid to prepare beauvericin. According to the method disclosed by the invention, the beauvericin is produced through liquid fermentation, and the prepared beauvericin is high in purity and recovery rate and adaptable in industrial production.

Description

technical field [0001] The invention belongs to the technical field of industrial microorganisms, and in particular relates to a preparation method of beauveria. Background technique [0002] Beauvericin (BEA) is derived from the fungus Beauvericin ( Beauverlabassiana ) a six-membered cyclic desipeptide compound extracted and isolated from fermented mycelia, consisting of 3 N-methylphenylalanine and 3 (D)- α -Hydroxyisovaleric acid residues are alternately connected, and the molecular formula is C 45 h 57 N 3 o 9 , the structure is shown in formula (I). Beauveriacin is in the form of white powder, not easily soluble in water, but soluble in organic solvents such as methanol and acetone. The detectable ultraviolet absorption spectrum ranges from 204nm to 225nm. [0003] [0004] Beauverum is a lipopeptide compound with biological activities such as antibacterial, insecticidal, antiviral, anticancer, inhibition of cholesterol acyltransferase activity, and induction of...

Claims

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Application Information

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IPC IPC(8): C12P21/04C07K1/16C12R1/77
Inventor 段宝玲张立新张雪霞刘梅任风芝贺伟李宁白超弦李晓露贾啸静陈书红米贯东李丽红王秀琴董爱华王海燕段向东
Owner NCPC NEW DRUG RES & DEV
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